Molecular cloning of a novel potassium-dependent sodium-calcium exchanger from rat brain.
We have isolated a novel cDNA clone from rat cerebral cortex encoding a protein of 670 amino acids (NCKX2) that has significant similarity to the 1199-amino acid-long Na/Ca-K exchanger of bovine rod outer segment (NCKX1). NCKX2 transcripts are 10.5 kilobase pairs in length and are expressed abundantly in neurons throughout the brain and with much lower abundance in selected other tissues. The predicted topology of the rat NCKX2 protein is very similar to that of bovine NCKX1, beginning with a solitary transmembrane segment (M0), which is removed as a "signal peptide" in bovine NCKX1, an extracellular loop, a cluster of five transmembrane spanning segments (M1 to M5), a long cytoplasmic loop, and a final hydrophobic cluster (M6 to M11). Within the hydrophobic clusters, rat NCKX2 shares 80% identity and 91% similarity with bovine NCKX1. The two larger hydrophilic loops are much shorter in NCKX2 than in NCKX1, accounting largely for the difference in length between the two proteins, and are dissimilar in sequence except for a 32-amino acid stretch with 69% identity in the cytosolic loop. NCKX2 was epitope-tagged in the extracellular domain and was shown to be expressed at the surface of transfected HEK cells. Analysis of NCKX2 function by fluorescent imaging of fura-2-loaded transfected cells demonstrated that NCKX2 is a potassium-dependent sodium/calcium exchanger.
Tsoi
M
M
Department of Medical Biochemistry, University of Calgary, Calgary, Alberta T2N 4N1, Canada.
Rhee
K H
KH
Bungard
D
D
Li
X F
XF
Lee
S L
SL
Auer
R N
RN
Lytton
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry*; D002417:Cattle; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D017403:In Situ Hybridization; D008565:Membrane Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry; D011188:Potassium / Q000494:pharmacology; D012333:RNA, Messenger; D051381:Rats; D017948:Retinal Rod Photoreceptor Cells / Q000737:chemistry; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D019831:Sodium-Calcium Exchanger / Q000737:chemistry*
D002352:Carrier Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D019831:Sodium-Calcium Exchanger; C110374:sodium-calcium exchanger 1; C074530:potassium-dependent sodium-calcium exchanger; D011188:Potassium; D002118:Calcium